Producing progeny from endangered birds of prey: treatment of urine-contaminated semen and a novel intramagnal insemination approach.

Wild raptors brought into an ex situ environment often have poor semen quality that is further compromised by urine contamination. Generally, it is believed that in birds, artificial insemination into the cloaca or caudal vagina of females requires large doses of high-quality spermatozoa to maximize fertility. In an effort to define and overcome some of the challenges associated with reproduction in wild raptors, the objectives of this study were to 1) evaluate the frequency, impact, and remediation of urine contamination in fresh ejaculates for the purpose of maintaining sperm motility and viability in vitro, and 2) develop a deep insemination method that allows low numbers of washed sperm to be placed directly into the magnum to increase the probability of producing fertilized eggs. The species evaluated include golden eagle (Aquila chrysoetos), imperial eagle (A. adalberti), Bonelli's eagle (Hiernaetus fasciatus), and peregrine falcon (Falco peregrinus). Semen samples were collected and pooled by species, and a minimum of 25 pooled ejaculates per species were evaluated for urine contamination, pH, sperm viability, and sperm motility; the samples were either unwashed or washed in neutral (pH 7.0) or alkaline (pH 8.0) modified Lake's diluent. Female golden eagles and peregrine falcons were inseminated via transjunctional, intramagnal insemination with washed spermatozoa from urine-contaminated samples. Urine contamination occurred in 36.8 +/- 12.8% (mean +/- SEM) golden eagle, 43.1 +/- 9.1% imperial eagle. 28.7 +/- 16.1% Bonelli's eagle, and 48.2 +/- 17.3% peregrine falcon ejaculates. The pH in urine-contaminated semen samples ranged from 6.48 +/- 0.3 to 6.86 +/- 0.2, and in noncontaminated samples it ranged from from 7.17 +/- 0.1 to 7.56 +/- 0.1. Sperm viability and motility were reduced (P < 0.05) in all species for unwashed vs. washed sperm after 30 min incubation at room temperature. Two peregrine falcon chicks and one golden eagle chick hatched after intramagnal insemination. This study demonstrates that urine contamination, a common and lethal acidifier in manually collected raptor ejaculates, can be circumvented by immediate, gentle seminal washing. Furthermore, these processed sperm, when deposited by transjunctional intramagnal insemination, can produce live young.     

CRANIAL MEDIASTINAL CYSTS IN NINE CATS

Nine cats, from 11 to 17 years of age (mean 13.6 years of age), were diagnosed with a cranial mediastinal cyst. Thoracic radiographs in all cats were characterized by an increased soft tissue opacity in the cranial mediastinum confirmed to be a cyst by ultrasonography or necropsy. Ultrasonographically cysts appeared as an anechoic mass. A low-cellularity clear fluid was obtained on aspiration. The majority of the cats (n = 8) presented for unrelated conditions with no signs of respiratory distress. No treatment for the cyst was pursued except for drainage during ultrasonographic-guided aspiration in several cats. On follow-up of eight cats, none were symptomatic for the cyst from 3-45 months after diagnosis. Mediastinal cyst should be considered when a cranial mediastinal mass is evident radiographically in an older cat. The majority of feline cranial mediastinal cysts are benign with no need for treatment.     

Isolation and characterization of bone marrow-derived equine mesenchymal stem cells

OBJECTIVE: To isolate and characterize bone marrow-derived equine mesenchymal stem cells (MSCs) for possible future therapeutic applications in horses. SAMPLE POPULATION: Equine MSCs were isolated from bone marrow aspirates obtained from the sternum of 30 donor horses. PROCEDURES: Cells were cultured in medium (alpha-minimum essential medium) with a fetal calf serum content of 20%. Equine MSC features were analyzed to determine selfrenewing and differentiation capacity. For potential therapeutic applications, the migratory potential of equine MSCs was determined. An adenoviral vector was used to determine the transduction rate of equine MSCs. RESULTS: Equine MSCs can be culture-expanded. Equine MSCs undergo cryopreservation in liquid nitrogen without altering morphologic characteristics. Furthermore, equine MSCs maintain their ability to proliferate and differentiate after thawing. Immunocytochemically, the expression of the stem cell marker CD90 can be detected on equine MSCs. The multilineage differentiation potential of equine MSCs was revealed by their ability to undergo adipogenic, osteogenic, and chondrogenic differentiation. CONCLUSIONS AND CLINICAL RELEVANCE: Our data indicate that bone marrow-derived stromal cells of horses can be characterized as MSCs. Equine MSCs have a high transduction rate and migratory potential and adapt to scaffold material in culture. As an autologous cell population, equine MSCs can be regarded as a promising cell population for tissue engineering in lesions of the musculoskeletal system in horses.     

Women

In New Zealand while women dominate leadership positions in constitutional, political and business areas, gender inequality remains entrenched. Despite significant gains in rights (employment, pay equity, property), as a group women earn less and perform more family care and unpaid work than men. In part these disparities result from the economic restructuring of the last 15 years which saw a loss in unskilled manufacturing jobs, less social support and greater rewards to business and public sector elites. Maori and Pacific Island women are the most disadvantaged groups. Labour/Alliance government initiatives to reduce poverty particularly among Maori and Pacific Island peoples by strengthening access to health and education services and supporting and replicating successful community initiatives are also likely to benefit other low income women.     

Effect of high-pressure hot-water washing treatment on fruit quality, insects, and disease in apples and pears: Part I. System description and the effect on fruit quality of ‘d’Anjou’ pears

A manually operated high-pressure hot-water washing system consisting of a boiler, hot-water mixing tank, contact loop, heat exchanger, spray mixing tank, high-pressure hot-water washing manifold, low-pressure fresh water rinse manifold, and pressure pump was constructed and installed in a packingline. The system developed 20–50 °C washing water at pressures up to 980 kPa. ‘d’Anjou’ pears (Pyrus communis L.), shortly after harvest, and after storage for 3 and 4 months in regular air (RA) or for 4, 7 and 8 months in controlled atmosphere (CA) at −1 °C were washed through the packingline with different wetting agents (0.1% Silwet, 0.01 and 0.1% Defoamer, and water), water pressures (regular and high-pressure (210–980 kPa)), water temperatures (control (tap water, 4–22 °C), 40 °C, and 50 °C), and brushes (soft and firm), respectively. The effect of the washing conditions on fruit quality was investigated after 1 month of storage at −1 °C to simulate shipping condition, and then again after 1 week at 20 °C to simulate marketing condition. Hot-water caused severe heat scald. When nozzle temperature was 50 °C, the incidence of heat scald increased to over 50% for the fruit stored in RA for 3 months. Combined with hot-water, 540 kPa high-pressure washing increased the incidence of friction discoloration. There were lower incidences of friction discoloration and heat scald for fruit stored in CA for 7 months, in comparison to that in RA for 3 months. However, those fruit did not ripen properly as indicated by a high extractable juice content. Fruit washed at harvest had minor incidences of friction discoloration regardless of different brushes, water pressures, and wetting agents. Fruit washed after storages in either 4 months RA or 4 or 8 months CA suffered a high incidence of friction discoloration including scuffing symptoms and pressure marking. The firm brushes caused a higher incidence of friction discoloration mainly because of scuffing symptoms. However, no differences were found between different water pressures and wetting agents with respect to friction discoloration. Fruit stored for 4 months RA suffered 26–28% friction discoloration in comparison to 16–18% in CA stored fruit with firm brush washing. Extended CA storage increased friction discoloration even with soft brush washing. The results suggest that a washing system with high-pressure spray, <30 °C warm water, wetting agent Defoamer and rotating soft brushes were significantly effective in removing surface pests and decay control without causing internal or external damage of fruit.     

Phosphatase activity does not limit the microbial use of low molecular weight organic-P substrates in soil

Plant roots and soil microorganisms contain significant quantities of low molecular weight (MW) phosphorylated nucleosides and sugars. Consequently, upon death these can represent a significant input of organic-P to the soil. Some of these organic-P substrates must first be dephosphorylated by phosphatases before being assimilated by the soil microbial community while others can be taken up directly from soil solution. To determine whether sorption or phosphatase activity was limiting the bioavailability of low MW organic-P in soil we compared the microbial uptake and C mineralization of a range of ¹⁴C-labeled organic-P substrates [glucose-6-phosphate, adenosine monophosphate (AMP), adenosine diphosphate (ADP) and adenosine triphosphate (ATP)] to that of the parent compounds (adenosine and glucose). In a fertile grassland soil we showed that at low organic-P substrate concentrations (<0.5 mM) phosphatase activity did not limit microbial uptake or mineralization in comparison to their non-phosphorylated counterparts. However, at high substrate concentrations (1-10 mM) the mineralization of the organic-P compounds was significantly lower than that of the non-phosphorylated compounds suggesting that phosphatase activity or microbial transporter capacity limited bioavailability. Sorption to the solid phase followed the series glucose<adenosine<G-6-P<AMP<ADP=ATP. However, sorption of the organic-P compounds to the solid phase did not appear to greatly affect bioavailability. The high adenosine mineralization capacity of the microbial biomass suggests that nucleosides may represent a significant source of C and N to the soil microbial biomass. We conclude that at low organic-P substrate concentrations typical of those in soil, neither phosphatase activity nor sorption greatly limits their bioavailability.     

Stress related changes in the architecture of seedlings of three provenances ofEucalyptus camaldulensis Dehnh

Abscisic acid (10^?4M) applied via absorbent discs bound to the stems of 6 wk oldE. camaldulensis seedlings caused changes in leaf shape and orientation similar to those induced by nutritional and water stress. Seedlings from the driest provenance, Tennant Creek, were more responsive to ABA than seedlings from the wettest provenance, Katherine, and the response of Petford seedlings was intermediate. Likewise, Tennant Creek seedlings were most, and Katherine seedlings least, responsive to water and/or nutritional stress. Seedlings that responded to the application of ABA produced prematurely strengthened, linear leaves with the lamina rotated from horizontal to vertical. The ability of Tennant Creek seedlings to readily assume this form may be a specific adaptation for the particularly difficult conditions in their natural environment.     

Changes in fruit antioxidant activity among blueberry cultivars during cold-temperature storage

Antioxidant activity, total phenolic content, anthocyanin content, and six other fruit characters including titratable acid concentration, soluble solids, firmness, and percentage of bruised berries were determined for nine blueberry (Vaccinium L. sp.) cultivars at harvest and at various postharvest intervals after storage at 5 degrees C. Berries from MSU-58, Brigitta, and Legacy stored successfully for 7 weeks, Bluegold stored for 3-5 weeks, Bluecrop, Elliott, and Nelson stored for 3 weeks, and Jersey and Little Giant stored for fewer than 3 weeks. During the time they retained marketable quality, one cultivar (MSU-58) demonstrated a 29% increase in antioxidant activity. None of the cultivars showed a significant decrease from the harvest antioxidant activity value during storage. Antioxidant activity, total phenolic content, and anthocyanin content were strongly correlated with each other (r = 0.87-0.99, P < 0.01). All three parameters were moderately correlated with soluble solids (r = 0.47, P < or =0.05; r = 0.44, P < or = 0.05; and r = 0.64, P < or = 0.01, respectively), and antioxidant activity and total phenolic content were both moderately correlated with pH (r = 0.53 and 0.49, respectively; P < or = 0.05). However, antioxidant activity, total phenolic content, and anthocyanin content showed no correlation with firmness, percent severely bruised berries, or weight loss. Antioxidant activity and total phenolic content at harvest both correlated with titratable acidity at harvest (r = 0.68, P < or = 0.05 and r = 0.70, P < or = 0.05, respectively) on a cultivar mean basis. Berries from Elliott were also harvested from plants at two levels of bush ripeness (30-50% and 60-80% ripe berries on plants) and separated into three fruit maturity classes on the basis of percent blue color. The level of bush ripeness had no significant effect on antioxidant activity, total phenolic content, or anthocyanin content; however, fruit maturity had a significant effect on antioxidant activity, total phenolic content, and anthocyanin content, and bush ripeness x fruit maturity interactions were significant for these three traits. Berries with 50-75% blue coloration harvested from bushes with 60-80% mature fruit showed a significant increase in antioxidant activity, total phenolic content, and anthocyanin content during the first 3 weeks in storage. Our results demonstrate that increases in antioxidant activity, total phenolic content, and anthocyanin content may occur in the blueberry during cold storage and are cultivar-dependent. The increases that occur in immature fruit, such as in Elliott, may be advantageous for producers who wish to delay marketing of the fruit.